Salmeterol restores secretory functions in cystic fibrosis airway submucosal gland serous cells.
The activity of the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) can be mediated by surface G protein-coupled receptors such as the beta(2)-adrenergic receptor. In this study, we explored the effect of a long-acting beta(2)-adrenergic agonist, salmeterol, on the CFTR-dependent secretory capacity of a human CF tracheal gland serous cell line (CF-KM4), homozygous for the delF508 mutation. We showed that, compared with the untreated CF serous cells, a 24-hour pre-incubation period with 200 nM salmeterol induced an 83% increase in delF508-CFTR-mediated chloride efflux. The restoration of the bioelectric properties is associated with increased apical surface pool of delF508-CFTR. Salmeterol induced a decrease in ion concentration and an increase in the level of hydration of the mucus packaged inside the CF secretory granules. The effects of salmeterol are not associated with a persistent production of cAMP. Western blotting on isolated secretory granules demonstrated immunoreactivity for CFTR and lysozyme. In parallel, we measured by atomic force microscopy an increased size of secretory granules isolated from CF serous cells compared with non-CF serous cells (MM39 cell line) and showed that salmeterol was able to restore a CF cell granule size similar to that of non-CF cells. To demonstrate that the salmeterol effect was a CFTR-dependent mechanism, we showed that the incubation of salmeterol-treated CF serous cells with CFTR-inh172 suppressed the restoration of normal secretory functions. The capacity of salmeterol to restore the secretory capacity of glandular serous cells suggests that it could also improve the airway mucociliary clearance in patients with CF.
Delavoie
Franck
F
INSERM UMRS 926, Laboratoire de Microscopie Electronique Analytique, 21 rue Clément Ader, B.P. 138, 51685 Reims Cedex 2, France. franck.delavoie@univ-reims.fr
Molinari
Michael
M
Milliot
Magali
M
Zahm
Jean-Marie
JM
Coraux
Christelle
C
Michel
Jean
J
Balossier
Gérard
G
D000420:Albuterol / Q000031:analogs & derivatives* / Q000494:pharmacology; D002460:Cell Line; D016764:Cell Polarity / Q000187:drug effects; D002712:Chlorides / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000096:biosynthesis; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D055724:Electrophysiological Phenomena / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005089:Exocytosis / Q000187:drug effects; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000648:ultrastructure; D007477:Ions / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D000068299:Salmeterol Xinafoate; D022142:Secretory Vesicles / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D012704:Serous Membrane / Q000378:metabolism* / Q000473:pathology*; D014132:Trachea / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology*
C505032:CFTR protein, human; D002712:Chlorides; D007477:Ions; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005576:Colforsin; D000068299:Salmeterol Xinafoate; D000242:Cyclic AMP; D009113:Muramidase; D000420:Albuterol